Blueberries Abstracts 2

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Nutraceutical interventions for promoting healthy aging in invertebrate models.
            (Dong et al., 2012) Download
Aging is a complex and inevitable biological process that is associated with numerous chronically debilitating health effects. Development of effective interventions for promoting healthy aging is an active but challenging area of research. Mechanistic studies in various model organisms, noticeably two invertebrates, Caenorhabditis elegans and Drosophila melanogaster, have identified many genes and pathways as well as dietary interventions that modulate lifespan and healthspan. These studies have shed light on some of the mechanisms involved in aging processes and provide valuable guidance for developing efficacious aging interventions. Nutraceuticals made from various plants contain a significant amount of phytochemicals with diverse biological activities. Phytochemicals can modulate many signaling pathways that exert numerous health benefits, such as reducing cancer incidence and inflammation, and promoting healthy aging. In this paper, we outline the current progress in aging intervention studies using nutraceuticals from an evolutionary perspective in invertebrate models.

Blueberry juice causes potent relaxation of rat aortic rings via the activation of potassium channels and the H₂S pathway.
            (Horrigan et al., 2013) Download
The objective of this study was to investigate the in vitro effects of blueberry juice on healthy rat aortic rings, and to explore the roles of potassium channels and of the hydrogen sulphide (H(2)S) pathway in mediating the effects of blueberry juice. Firstly, the antioxidant capacity of blueberry juice was compared to other popular juice drinks using the Folin-Ciocalteu and the DPPH assays. Blueberry juice had significantly higher total polyphenol content than any of the other drinks studied (p < 0.01). The effect of blueberry juice on noradrenaline-contracted aortic rings was then observed, and the juice caused significant inhibition of noradrenaline-induced contractions (p < 0.01). Voltage-gated potassium channel (Kv) blockers 4-aminopyridine (1 mM) and 3,4-diaminopyridine (1 mM), as well as the cystathionine γ-lysase (CSE) inhibitor d,l-propargylglycine (2 mM) were then utilised to elucidate the role of Kv channels and the CSE/H(2)S pathway. Kv channel blocker 3,4-diaminopyridine caused significant blockade at 1/100 and 1/50 dilutions of juice (p < 0.01), whilst 4-aminopyridine caused significant blockade of the 1/100 dilution of blueberry juice (p < 0.05). In addition, d,l-propargylglycine potently inhibited the effect of 1/100 and 1/50 dilutions of blueberry juice (p < 0.01). This study indicates that blueberry juice has potent vasorelaxing properties, and thus may be a useful dietary agent for the prevention and treatment of hypertension. This study also provides strong evidence that Kv channels and the CSE/H(2)S pathway may be responsible, at least in part, for mediating the effects of blueberry juice.

Vascular reactivity is affected by dietary consumption of wild blueberries in the Sprague-Dawley rat.
            (Kalea et al., 2009) Download
We have previously reported that consumption of blueberry-enriched (BB) diets attenuates the arterial contractile response to alpha(1)-adrenergic stimuli and affects vasomotor tone via endothelium-related pathways. The present study was designed to evaluate vascular function and responsiveness in aortas of weanling male Sprague-Dawley rats fed a control (C) or a BB diet for 7 weeks. Vascular ring studies were conducted in 3-mm isolated rat aortic ring preparations to investigate vasoconstriction induced by L-phenylephrine (Phe) (10(-8)-3 x 10(-6) M) and vasorelaxation induced by acetylcholine (ACh) (10(-8)-3 x 10(-6) M). Agonists were used alone and in the presence of nitric oxide (NO) synthase and cyclooxygenase (COX) inhibitors. We observed a significantly diminished vasoconstrictor response to Phe in aortic rings from rats fed the BB diet. Inhibition of NO synthase but not COX caused a significant increase in the constrictor response in both dietary groups, with the BB group having the greater response. Similarly, the participation of the NO pathway in endothelium-dependent vasorelaxation induced by ACh was greater in the rats fed a BB diet, while COX inhibition showed no effect on maximum ACh-induced vasorelaxation in any diet group. The vessel sensitivity of BB aortic rings to the vasoconstrictor and vasodilator was significantly reduced when compared to controls. We have concluded that diets enriched with blueberries, fed for 7 weeks in Sprague-Dawley rats, seem to affect NO metabolic pathways in the aorta at basal and stimulated levels.

Cytosolic protection against ultraviolet induced DNA damage by blueberry anthocyanins and anthocyanidins in hepatocarcinoma HepG2 cells.
            (Liu et al., 2013) Download
UV-induced DNA damage plays a key role in the etiology of certain diseases. The ability of blueberry anthocyanins and anthocyanidins (BA) to protect cellular DNA from UV-induced damage was investigated. BA were extracted by water (BAW), ethanol (BAE) or methanol (BAM). These extracts partially restored proliferation of UV-irradiated HepG2 cells as shown by MTT assay. Treatment with BA extracts at 75 μg/ml decreased reactive oxygen species and decreased DNA damage by tail moment of comet assay and expression of γH2AX in situ. BAM significantly decreased gene and protein expression of p53, phospho-p53 (Ser15), and p21 in UV-irradiated HepG2 cells. BA thus efficiently protects cells from DNA damage in vitro. Blueberry may potentially be used as a good source of naturally radioprotective agents.

Blueberry extract prolongs lifespan of Drosophila melanogaster.
            (Peng et al., 2012) Download
Blueberry possesses greater antioxidant capacity than most other fruits and vegetables. The present study investigated the lifespan-prolonging activity of blueberry extracts in fruit flies and explored its underlying mechanism. Results revealed that blueberry extracts at 5mg/ml in diet could significantly extend the mean lifespan of fruit flies by 10%, accompanied by up-regulating gene expression of superoxide dismutase (SOD), catalase (CAT) and Rpn11 and down-regulating Methuselah (MTH) gene. Intensive H(2)O(2) and Paraquat challenge tests showed that lifespan was only extended in Oregon-R wild type flies but not in SOD(n108) or Cat(n1) mutant strains. Chronic Paraquat exposure shortened the maximum survival time from 73 to 35days and decreased the climbing ability by 60% while blueberry extracts at 5mg/ml in diet could significantly increase the survival rate and partially restore the climbing ability with up-regulating SOD, CAT, and Rpn11. Furthermore, gustatory assay demonstrated that those changes were not due to the variation of food intake between the control and the experimental diet containing 5mg/ml blueberry extracts. It was therefore concluded that the lifespan-prolonging activity of blueberry extracts was at least partially associated with its interactions with MTH, Rpn11, and endogenous antioxidant enzymes SOD and CAT.

Purified blueberry anthocyanins and blueberry juice alter development of obesity in mice fed an obesogenic high-fat diet.
            (Prior et al., 2010) Download
Male C57BL/6J mice (25 days of age) were fed either a low-fat diet (10% kcal from fat) (LF) or a high-fat diet (45% kcal from fat) (HF45) for a period of 72 days. Blueberry juice or purified blueberry anthocyanins (0.2 or 1.0 mg/mL) in the drinking water were included in LF or HF45 treatments. Sucrose was added to the drinking water of one treatment to test if the sugars in blueberry juice would affect development of obesity. Total body weights (g) and body fat (%) were higher and body lean tissue (%) was lower in the HF45 fed mice compared to the LF fed mice after 72 days, but in mice fed HF45 diet plus blueberry juice or blueberry anthocyanins (0.2 mg/mL), body fat (%) was not different from those mice fed the LF diet. Anthocyanins (ACNs) decreased retroperitoneal and epididymal adipose tissue weights. Fasting serum glucose concentrations were higher in mice fed the HF45 diet. However, it was reduced to LF levels in mice fed the HF45 diet plus 0.2 mg of ACNs/mL in the drinking water, but not with blueberry juice. beta cell function (HOMA-BCF) score was lowered with HF45 feeding but returned to normal levels in mice fed the HF45 diet plus purified ACNs (0.2 mg/mL). Serum leptin was elevated in mice fed HF45 diet, and feeding either blueberry juice or purified ACNs (0.2 mg/mL) decreased serum leptin levels relative to HF45 control. Sucrose in drinking water, when consumption was restricted to the volume of juice consumed, produced lower serum leptin and insulin levels, leptin/fat, and retroperitoneal and total fat (% BW). Blueberry juice was not as effective as the low dose of anthocyanins in the drinking water in preventing obesity. Additional studies are needed to determine factors responsible for the differing responses of blueberry juice and whole blueberry in preventing the development of obesity.

Blueberry intervention improves vascular reactivity and lowers blood pressure in high-fat-, high-cholesterol-fed rats.
            (Rodriguez-Mateos et al., 2013) Download
Growing evidence suggests that intake of flavonoid-containing foods may exert cardiovascular benefits in human subjects. We have investigated the effects of a 10-week blueberry (BB) supplementation on blood pressure (BP) and vascular reactivity in rats fed a high-fat/high-cholesterol diet, known to induce endothelial dysfunction. Rats were randomly assigned to follow a control chow diet, a chow diet supplemented with 2 % (w/w) BB, a high-fat diet (10 % lard; 0·5 % cholesterol) or the high fat plus BB for 10 weeks. Rats supplemented with BB showed significant reductions in systolic BP (SBP) of 11 and 14 %, at weeks 8 and 10, respectively, relative to rats fed the control chow diet (week 8 SBP: 107·5 (SEM 4·7) v. 122·2 (SEM 2·1) mmHg, P= 0·018; week 10 SBP: 115·0 (SEM 3·1) v. 132·7 (SEM 1·5) mmHg, P< 0·0001). Furthermore, SBP was reduced by 14 % in rats fed with the high fat plus 2 % BB diet at week 10, compared to those on the high-fat diet only (SBP: 118·2 (SEM 3·6) v. 139·5 (SEM 4·5) mmHg, P< 0·0001). Aortas harvested from BB-fed animals exhibited significantly reduced contractile responses (to L-phenylephrine) compared to those fed the control chow or high-fat diets. Furthermore, in rats fed with high fat supplemented with BB, aorta relaxation was significantly greater in response to acetylcholine compared to animals fed with the fat diet. These data suggest that BB consumption can lower BP and improve endothelial dysfunction induced by a high fat, high cholesterol containing diet.

Blueberry polyphenol-enriched soybean flour reduces hyperglycemia, body weight gain and serum cholesterol in mice.
            (Roopchand et al., 2013) Download
Defatted soybean flour (DSF) can sorb and concentrate blueberry anthocyanins and other polyphenols, but not sugars. In this study blueberry polyphenol-enriched DSF (BB-DSF) or DSF were incorporated into very high fat diet (VHFD) formulations and provided ad libitum to obese and hyperglycemic C57BL/6 mice for 13 weeks to investigate anti-diabetic effects. Compared to the VHFD containing DSF, the diet supplemented with BB-DSF reduced weight gain by 5.6%, improved glucose tolerance, and lowered fasting blood glucose levels in mice within 7 weeks of intervention. Serum cholesterol of mice consuming the BB-DSF-supplemented diet was 13.2% lower than mice on the diet containing DSF. Compounds were eluted from DSF and BB-DSF for in vitro assays of glucose production and uptake. Compared to untreated control, doses of BB-DSF eluate containing 0.05-10μg/μL of blueberry anthocyanins significantly reduced glucose production by 24-74% in H4IIE rat hepatocytes, but did not increase glucose uptake in L6 myotubes. The results indicate that delivery of blueberry polyphenols stabilized in a high-protein food matrix may be useful for the dietary management of pre-diabetes and/or diabetes.

Blueberry intake alters skeletal muscle and adipose tissue peroxisome proliferator-activated receptor activity and reduces insulin resistance in obese rats.
            (Seymour et al., 2011) Download
Metabolic syndrome can precede the development of type 2 diabetes and cardiovascular disease and includes phenotypes such as obesity, systemic inflammation, insulin resistance, and hyperlipidemia. A recent epidemiological study indicated that blueberry intake reduced cardiovascular mortality in humans, but the possible genetic mechanisms of this effect are unknown. Blueberries are a rich source of anthocyanins, and anthocyanins can alter the activity of peroxisome proliferator-activated receptors (PPARs), which affect energy substrate metabolism. The effect of blueberry intake was assessed in obesity-prone rats. Zucker Fatty and Zucker Lean rats were fed a higher-fat diet (45% of kcal) or a lower-fat diet (10% of kcal) containing 2% (wt/wt) freeze-dried whole highbush blueberry powder or added sugars to match macronutrient and calorie content. In Zucker Fatty rats fed a high-fat diet, the addition of blueberry reduced triglycerides, fasting insulin, homeostasis model index of insulin resistance, and glucose area under the curve. Blueberry intake also reduced abdominal fat mass, increased adipose and skeletal muscle PPAR activity, and affected PPAR transcripts involved in fat oxidation and glucose uptake/oxidation. In Zucker Fatty rats fed a low-fat diet, the addition of blueberry also significantly reduced liver weight, body weight, and total fat mass. Finally, Zucker Lean rats fed blueberry had higher body weight and reduced triglycerides, but all other measures were unaffected. In conclusion, whole blueberry intake reduced phenotypes of metabolic syndrome in obesity-prone rats and affected PPAR gene transcripts in adipose and muscle tissue involved in fat and glucose metabolism.


 

Wild blueberry (Vaccinium angustifolium) consumption improves inflammatory status in the obese Zucker rat model of the metabolic syndrome.
            (Vendrame et al., 2013) Download
The metabolic syndrome (MetS) is a major public health problem in the United States. Chronic inflammation is a critical component of the MetS, leading to dramatically increased risk of type II diabetes and cardiovascular disease. This study investigates the ability of a wild-blueberry-enriched diet to improve the proinflammatory status associated with MetS in the obese Zucker rat (OZR). Circulating levels of key inflammatory markers and their expression in the liver and abdominal adipose tissue were examined in OZR and its genetic control, the lean Zucker rat (LZR), after feeding a control or an 8% wild blueberry diet (WB) for 8 weeks from age 8 to 16 weeks. In the OZR, WB consumption resulted in decreased plasma concentrations of tumor necrosis factor (TNF)-α (-25.6%, P<.05), interleukin (IL)-6 (-14.9%, P<.05) and C-reactive protein (CRP) (-13.1%, P<.05) and increased adiponectin concentration (+21.8%, P<.05). Furthermore, expression of IL-6, TNF-α and nuclear factor (NF)-kB was down-regulated in both the liver (-65%, -59% and -25%, respectively) and the abdominal adipose tissue (-64%, -52% and -65%), while CRP expression was down-regulated only in the liver (-25%). In the abdominal adipose tissue, similar trends were also observed in LZR following WB treatment, with decreased liver expression of NF-kB, CRP, IL-6 and TNF-α (-24%, -16%, -21% and -50%) and increased adiponectin expression (+25%). Results of this study suggest that wild blueberry consumption exerts an overall anti-inflammatory effect in the OZR, a model of the metabolic syndrome.

Wild blueberry consumption affects aortic vascular function in the obese Zucker rat.
            (Vendrame et al., 2014) Download
This study evaluates the effect of wild blueberry (WB) consumption on the biomechanical properties of the aorta in the obese Zucker rat (OZR), a model of the metabolic syndrome. Thirty-six OZRs and 36 lean controls (lean Zucker rats) were placed either on a WB-enriched or a control (C) diet for 8 weeks. Phenylephrine (Phe)-mediated vasoconstriction and acetylcholine (Ach)-mediated vasorelaxation in the aortic vessel were investigated, as well as the contribution of the nitric oxide synthase and cyclooxygenase (COX) pathways in each of the above responses by using specific inhibitors. Obese Zucker rats exhibited a reduced vasocontstrictor response to Phe and an exaggerated vasorelaxant response to Ach. The WB diet partially restored Phe-induced constrictor responses and attenuated Ach-induced relaxant responses in OZR. Plasma nitric oxide was significantly attenuated (22.1 ± 1.1 μmol·L(-1), WB vs 25.6 ± 1.4 μmol·L(-1), C, p ≤ 0.05) with the WB diet. Thromboxane A2 levels in the aortic effluent were not significantly affected in the WB diet group, while PGI2 concentration significantly increased (766.5 ± 92.2 pg·mg(-1) aorta in the WB vs 571.7 ± 37.8 pg·g(-1) aorta in the C group, p ≤ 0.05). Downregulation of inducible nitric oxide synthase and COX2 expression in the OZR aorta was observed in the WB diet group. In conclusion, WB consumption altered the biomechanical properties of the OZR aorta by partially restoring the impaired Phe-induced constrictor responses and attenuating the exaggerated response to Ach-induced vasorelaxation.

 


References

Dong, Y, et al. (2012), ‘Nutraceutical interventions for promoting healthy aging in invertebrate models.’, Oxid Med Cell Longev, 2012 718491. PubMed: 22991584
Horrigan, LA, et al. (2013), ‘Blueberry juice causes potent relaxation of rat aortic rings via the activation of potassium channels and the H₂S pathway.’, Food Funct, 4 (3), 392-400. PubMed: 23175156
Kalea, AZ, et al. (2009), ‘Vascular reactivity is affected by dietary consumption of wild blueberries in the Sprague-Dawley rat.’, J Med Food, 12 (1), 21-28. PubMed: 19298192
Liu, W, et al. (2013), ‘Cytosolic protection against ultraviolet induced DNA damage by blueberry anthocyanins and anthocyanidins in hepatocarcinoma HepG2 cells.’, Biotechnol Lett, 35 (4), 491-98. PubMed: 23192380
Peng, C, et al. (2012), ‘Blueberry extract prolongs lifespan of Drosophila melanogaster.’, Exp Gerontol, 47 (2), 170-78. PubMed: 22197903
Prior, RL, et al. (2010), ‘Purified blueberry anthocyanins and blueberry juice alter development of obesity in mice fed an obesogenic high-fat diet.’, J Agric Food Chem, 58 (7), 3970-76. PubMed: 20148514
Rodriguez-Mateos, A, et al. (2013), ‘Blueberry intervention improves vascular reactivity and lowers blood pressure in high-fat-, high-cholesterol-fed rats.’, Br J Nutr, 109 (10), 1746-54. PubMed: 23046999
Roopchand, DE, et al. (2013), ‘Blueberry polyphenol-enriched soybean flour reduces hyperglycemia, body weight gain and serum cholesterol in mice.’, Pharmacol Res, 68 (1), 59-67. PubMed: 23220243
Seymour, EM, et al. (2011), ‘Blueberry intake alters skeletal muscle and adipose tissue peroxisome proliferator-activated receptor activity and reduces insulin resistance in obese rats.’, J Med Food, 14 (12), 1511-18. PubMed: 21861718
Vendrame, S, et al. (2013), ‘Wild blueberry (Vaccinium angustifolium) consumption improves inflammatory status in the obese Zucker rat model of the metabolic syndrome.’, J Nutr Biochem, 24 (8), 1508-12. PubMed: 23465589
Vendrame, S, et al. (2014), ‘Wild blueberry consumption affects aortic vascular function in the obese Zucker rat.’, Appl Physiol Nutr Metab, 39 (2), 255-61. PubMed: 24476483